Rapid evaluation of the efficacy of microbial cell removal from fabrics

  • Kohtaro Fujioka
  • Ikuko Kozone
  • Mikako Saito
  • Hideaki MatsuokaEmail author
Original Paper


The efficacy of microbial cell removal (EMR) from fabrics is a practically important indicator for the evaluation of cleansers and detergents. EMR is expressed quantitatively by the relative number of viable cells remaining on a fabric swatch after the treatment with these reagents. In order to count the viable cells on the swatch directly and rapidly, we have developed a unique microscopic imaging system with an ultra-deep focusing range. Standard swatches of cotton fabric were inoculated with microorganisms such as Pseudomonas fluorescence, Staphylococcus aureus, or Candida albicans. After the incubation on an agar medium, each swatch was treated with a fluorescent glucose, 2-[N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino]-2-deoxyglucose, to stain only viable cells. The images of every cell distributed within the surface layer with no greater than 130 μm thickness could be integrated into one image. Thus visualized cells could be counted automatically by a novel imaging program. Using a pair of cotton swatches (0.5×1.0 cm2) inoculated with C. albicans, EMR was evaluated quantitatively. Before washing, the total number of viable cells found on the observation area (3.8×10−4 cm2) was 288 cells. After washing with a test detergent, no cell (<1) was detected. For this case, EMR was given by the formula: log(288/<1)=greater than 2.5. The imaging and cell count of a test fabric could be performed within 1 h.


Ultra-deep focusing range (UDF) fluorescent microscope Efficacy of microbial cell removal (EMR) A fluorescent glucose Viable cell imaging 



This research was supported by the Microbial Visualization Community of Practice in Procter & Gamble Company. The authors would like to thank Dr. P. Geis and Dr. S. Donaldson in Procter & Gamble Company for the constructive discussions and for reviewing the manuscript and Mr. Tottori of Kogaku Inc. for the supports and inputs for the microscopic system designs. This work is partially supported by the know-hows developed in the research project: The High Throughput Creation of Disease Model Cells and the Analysis of Their Function, which is funded by Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency.


  1. 1.
    ASTM E2274-03 (2004) Standard test method for evaluation of laundry sanitizers and disinfectantsGoogle Scholar
  2. 2.
    Borkow G, Gabbay J (2004) Putting copper into action: copper-impregnated products with potent biocidal activities. FASEB J 18:1728–1730Google Scholar
  3. 3.
    Brannan D-K, Dille J-C, Kaufman D-J (1987) Correlation of in vitro challenge testing with consumer use testing for cosmetic products. Appl Environ Microbiol 53:1827–1832Google Scholar
  4. 4.
    Buda A, Sands C, Jepson M-A (2005) Use of fluorescence imaging to investigate the structure and function of intestinal M cells. Adv Drug Deliv Rev 57:123–134CrossRefGoogle Scholar
  5. 5.
    Burton K (2003) An aperture-shifting light-microscopic method for rapidly quantifying positions of cells in 3D matrices. Cytometry A 54:125–131CrossRefGoogle Scholar
  6. 6.
    Cen L, Neoh K-G, Kang E-T (2004) Antibacterial activity of cloth functionalized with N-alkylated poly(4-vinylpyridine). J Biomed Mater Res A 71:70–80CrossRefGoogle Scholar
  7. 7.
    JIS L0803 (1998) Standard adjacent fabrics for straining of colour fastness testGoogle Scholar
  8. 8.
    JIS L0844 (1997) Test methods for colour fastness to washing and launderingGoogle Scholar
  9. 9.
    JIS L1902 (2002) Testing for antibacterial activity and efficacy on textile productsGoogle Scholar
  10. 10.
    Klueh U, Wagner V, Kelly S, Johnson A, Bryers J-D (2000) Efficacy of silver-coated fabric to prevent bacterial colonization and subsequent device-based biofilm formation. J Biomed Mater Res 53:621–631CrossRefGoogle Scholar
  11. 11.
    Laflamme C, Lavigne S, Ho J, Duchaine C. (2004) Assessment of bacterial endospore viability with fluorescent dyes. J Appl Microbiol 96:684–692CrossRefGoogle Scholar
  12. 12.
    Larson E-L, Lin S-X, Gomez-Pichardo C, Della-Latta P (2004) Effect of antibacterial home cleaning and handwashing products on infectious disease symptoms: a randomized, double-blind trial. Ann Intern Med 140(5):321–329Google Scholar
  13. 13.
    Lepeuple A-S, Gilouppe S, Pierlot E, de Roubin M-R. (2004) Rapid and automated detection of fluorescent total bacteria in water samples. Inter J Food Microbiol 92:327–332CrossRefGoogle Scholar
  14. 14.
    Lin J, Qiu S, Lewis K, Klibanov A-M (2003) Mechanism of bactericidal and fungicidal activities of textiles covalently modified with alkylated polyethylenimine. Biotechnol Bioeng 83:168–172CrossRefGoogle Scholar
  15. 15.
    Matsuoka H., Kurokawa T, Oishi K, Saito M (2002) Rapid detection of a small number of viable cells by the fluorescent glucose method. J Food Safety 22:141–153Google Scholar
  16. 16.
    Matsuoka H, Oishi K, Watanabe M, Kozone I, Saito M, Igimi S (2003) Viable cell detection by the combined use of fluorescent glucose and fluorescent glycine. Biosci Biotechnol Biochem 67:2459–2462CrossRefGoogle Scholar
  17. 17.
    McNally J-G, Karpova T, Cooper J, Conchello J-A (1999) Three-dimensional imaging by deconvolution microscopy. Methods 19:373–385CrossRefGoogle Scholar
  18. 18.
    Norman K (2005) Techniques: intravital microscopy—a method for investigating disseminated intravascular coagulation? Trends Pharmacol Sci 26:327–332CrossRefGoogle Scholar
  19. 19.
    Petrocci A-M, Clarke P (1969) Proposed test method for antimicrobial laundry additives. J AOAC 52:836–842Google Scholar
  20. 20.
    Roldán M, Thomas F, Castel S, Quesada A, Hernández-Mariné M (2004) Noninvasive pigment identification in single cells from living phototrophic biofilms by confocal imaging spectrofluorometry. Appl Env Microb 70:3745–3750CrossRefGoogle Scholar
  21. 21.
    Staudt C, Horn H, Hempel D-C, Neu T-R (2004) Volumetric measurements of bacterial cells and extracellular polymeric substance glycoconjugates in biofilms. Biotechnol Bioeng 88:585–592CrossRefGoogle Scholar
  22. 22.
    Sunamura M, Maruyama A, Tsuji T, Kurane R. (2003) Spectral imaging detection and counting of microbial cells in marine sediment. J Microbiol Method 53:57–65CrossRefGoogle Scholar
  23. 23.
    Takai K, Ohtsuka T, Senda Y, Nakao M, Yamamoto K, Matsuoka J, Hirai Y (2002) Antibacterial properties of antimicrobial-finished textile products. Microbiol Immunol 46:75–81Google Scholar
  24. 24.
    USEPA 712-C-97–056 (1997a) General requirements for public health uses of antimicrobial agents. Product Performance Test Guidelines OPPTS 810.2000Google Scholar
  25. 25.
    USEPA 712-C-97–056 (1997b) Products for use on hard surfaces – Basic efficacy data requirements. Product Performance Test Guidelines OPPTS 810.2100Google Scholar
  26. 26.
    USEPA 712-C-97–091 (1997c) Products for use on fabrics and textiles. Product Performance Test Guidelines OPPTS 810.2300Google Scholar
  27. 27.
    Vermeersch G, Leloup G, Delmee M, Vreven J (2005) Antibacterial activity of glass–ionomer cements, compomers and resin composites: relationship between acidity and material setting phase. J Oral Rehabil 32:368–374CrossRefGoogle Scholar
  28. 28.
    Yamada K, Nakata M, Horimoto N, Saito M, Matsuoka H, Inagaki N (2000) Measurement of glucose uptake and intracellular calcium concentration in single, living pancreatic β-cells. J Biol Chem 275:22278–22283CrossRefGoogle Scholar
  29. 29.
    Yoshioka K, Takahashi H, Homma T, Saito M, Oh K.-B, Nemoto Y, Matsuoka H (1996) A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli. Biochim Biophys Acta 1289:5–9Google Scholar

Copyright information

© Society for Industrial Microbiology 2006

Authors and Affiliations

  • Kohtaro Fujioka
    • 1
  • Ikuko Kozone
    • 2
    • 3
  • Mikako Saito
    • 2
    • 3
  • Hideaki Matsuoka
    • 2
    • 3
    Email author
  1. 1.Kobe Technical CenterProcter & Gamble Far East, Inc.KobeJapan
  2. 2.Department of Biotechnology and Life ScienceTokyo University of Agriculture and TechnologyTokyoJapan
  3. 3.CREST, Japan Science and Technology AgencySaitamaJapan

Personalised recommendations